Chromosome-level genome assembly reveals the genetic mechanisms underlying elite waterlogging tolerance in Actinidia rufa

Qiong Zhang , Linhui Wu , Hao Zheng , Yule Wang , Zupeng Wang , Shaokui Yi , Yifei Liu , Caihong Zhong

Horticulture Advances ›› 2026, Vol. 4 ›› Issue (1) : 14

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Horticulture Advances ›› 2026, Vol. 4 ›› Issue (1) :14 DOI: 10.1007/s44281-026-00103-z
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Chromosome-level genome assembly reveals the genetic mechanisms underlying elite waterlogging tolerance in Actinidia rufa
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Abstract

Actinidia rufa is a valuable species for hybridization and introgressive breeding of new kiwifruit cultivars due to its exceptional tolerance to waterlogging. In this study, a chromosome-level genome of A. rufa was assembled using Illumina short-read and PacBio Continuous Long Read (CLR) sequencing technologies. The assembled genome spans 613.66 Mb with an N50 of 3.06 Mb, and 94 scaffolds were anchored to 29 pseudochromosomes. A total of 42,484 protein-coding genes and 267.26 Mb of repetitive sequences, accounting for 49.08% of the genome, were identified. Phylogenetic analysis demonstrated a close evolutionary relationship between A. rufa and A. chinensis, with their most recent common ancestor being estimated to diverge around 7 million years ago. Demographic analysis indicated a historical population expansion in A. rufa, whose current suitable habitats are primarily found in the Japanese archipelago and Taiwan, China. Comparative genomic analysis revealed large-scale structural variations, including a chromosomal region on Chr19 in A. rufa that corresponds to two distinct segments on Chr2 and Chr9 in A. chinensis. Additionally, A. rufa cv. ‘MTS7001’ exhibited superior waterlogging tolerance compared to A. chinensis cv. ‘Donghong’. Gene expression analysis showed that, in response to early waterlogging stress, A. rufa exhibited upregulation of genes associated with energy metabolism and signaling of hormones, including those related to auxin, abscisic acid, and brassinosteroids, compared to A. chinensis. This study provides genomic insights into the genetic basis of exceptional waterlogging tolerance in A. rufa, contributing to molecular breeding efforts in kiwifruits in the future.

Keywords

Kiwifruit / Actinidia / Gene regulation / Waterlogging stress / RNA-seq

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Qiong Zhang, Linhui Wu, Hao Zheng, Yule Wang, Zupeng Wang, Shaokui Yi, Yifei Liu, Caihong Zhong. Chromosome-level genome assembly reveals the genetic mechanisms underlying elite waterlogging tolerance in Actinidia rufa. Horticulture Advances, 2026, 4(1): 14 DOI:10.1007/s44281-026-00103-z

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References

[1]

Adey A, Kitzman JO, Burton JN, Daza R, Kumar A, Christiansen L, et al.. In vitro, long-range sequence information for de novo genome assembly via transposase contiguity. Genome Res, 2014, 24: 2041-2049

[2]

Akagi T, Varkonyi-Gasic E, Shirasawa K, Catanach A, Henry IM, Mertten D, et al.. Recurrent neo-sex chromosome evolution in kiwifruit. Nat Plants, 2023, 9: 393-402

[3]

Alam I, Lee DG, Kim KH, Park CH, Sharmin SA, Lee H, et al.. Proteome analysis of soybean roots under waterlogging stress at an early vegetative stage. J Biosci, 2010, 35: 49-62

[4]

Alpuerto JB, Hussain RMF, Fukao T. The key regulator of submergence tolerance, SUB1A, promotes photosynthetic and metabolic recovery from submergence damage in rice leaves. Plant Cell Environ, 2016, 39: 672-684

[5]

Bao W, Kojima KK, Kohany O. Repbase update, a database of repetitive elements in eukaryotic genomes. Mob DNA, 2015, 6: 11

[6]

Benson G. Tandem repeats finder: a program to analyze DNA sequences. Nucleic Acids Res, 1999, 27: 573-580

[7]

Chen S. Ultrafast one-pass FASTQ data preprocessing, quality control, and deduplication using fastp. iMeta. 2023;2:e107. https://doi.org/10.1002/imt2.107.

[8]

Chen S, Zhou Y, Chen Y, et al.. Fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics, 2018, 34: i884-i890

[9]

Chin CS, Peluso P, Sedlazeck FJ, Nattestad M, Concepcion GT, Clum A, et al.. Phased diploid genome assembly with single-molecule real-time sequencing. Nat Methods, 2016, 13: 1050-1054

[10]

De Bie T, Cristianini N, Demuth JP, Hahn MW. CAFE: a computational tool for the study of gene family evolution. Bioinformatics, 2006, 22: 1269-1271

[11]

Dong B, Wang Q, Zhou D, Wang Y, Miao Y, Zhong S, et al.. Abiotic stress treatment reveals expansin like A gene OfEXLA1 improving salt and drought tolerance of Osmanthus fragrans by responding to abscisic acid. Hortic Plant J, 2024, 10: 573-585

[12]

Emms DM, Kelly S. OrthoFinder: phylogenetic orthology inference for comparative genomics. Genome Biol, 2019, 20: 238

[13]

Flynn JM, Hubley R, Goubert C, Rosen J, Clark AG, Feschotte C, et al.. RepeatModeler2 for automated genomic discovery of transposable element families. Proc Natl Acad Sci USA, 2020, 117: 9451-9457

[14]

Gao F, Chen C, Arab DA, Du Z, He Y, Ho SYW. EasyCodeML: a visual tool for analysis of selection using CodeML. Ecol Evol, 2019, 9: 3891-3898

[15]

Gao Y, Jiang Z, Shi M, Zhou Y, Huo L, Li X, et al.. Comparative transcriptome provides insight into responding mechanism of waterlogging stress in Actinidia valvata Dunn. Gene, 2022, 845: 146843

[16]

Geng S, Lin Z, Xie S, Xiao J, Wang H, Zhao X, et al.. Ethylene enhanced waterlogging tolerance by changing root architecture and inducing aerenchyma formation in maize seedlings. J Plant Physiol, 2023, 287: 154042

[17]

Gubitosa J, Rizzi V, Cignolo D, Fini P, Fanelli F, Cosma P. From agricultural wastes to a resource: kiwi peels, as long-lasting, recyclable adsorbent, to remove emerging pollutants from water. The case of ciprofloxacin removal. Sustain Chem Pharm, 2022, 29: 100749

[18]

Haas BJ, Papanicolaou A, Yassour M, Grabherr M, Blood PD, Bowden J, et al.. De novo transcript sequence reconstruction from RNA-seq using the Trinity platform for reference generation and analysis. Nat Protoc, 2013, 8: 1494-1512

[19]

Haas BJ, Salzberg SL, Zhu W, Perțea M, Allen JE, Orvis J, et al.. Automated eukaryotic gene structure annotation using EVidenceModeler and the Program to Assemble Spliced Alignments. Genome Biol, 2007, 9: R7

[20]

Han X, Zhang Y, Zhang Q, Ma N, Liu X, Tao W, et al.. Two haplotype-resolved, gap-free genome assemblies for Actinidia latifolia and Actinidia chinensis shed light on the regulatory mechanisms of vitamin C and sucrose metabolism in kiwifruit. Mol Plant, 2022, 16: 452-470

[21]

He W, Luo L, Xie R, Chai J, Wang H, Wang Y, et al.. Genome-wide identification and functional analysis of the AP2/ERF transcription factor family in citrus rootstock under waterlogging stress. Int J Mol Sci, 2023, 24: 8989

[22]

Huang H, Liu Y. Natural hybridization, introgression breeding, and cultivar improvement in the genus Actinidia. Tree Genet Genomes, 2014, 10: 1113-1122

[23]

Huang S, Ding J, Deng D, Tang W, Sun H, Liu D, et al.. Draft genome of the kiwifruit Actinidia chinensis. Nat Commun, 2013, 4 Article ID: 2640

[24]

Juntawong P, Sirikhachornkit A, Pimjan R, Sonthirod C, Sangsrakru D, Yoocha T, et al.. Elucidation of the molecular responses to waterlogging in Jatropha roots by transcriptome profiling. Front Plant Sci, 2014, 5: 658

[25]

Kim D, Pertea G, Trapnell C, Pimentel H, Kelley R, Salzberg SL. TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions. Genome Biol, 2013, 14: R36

[26]

Kisaki G, Tanaka S, Ishihara A, Igarashi C, Morimoto T, Hamano K, et al.. Evaluation of various cultivars of Actinidia species and breeding source Actinidia rufa for resistance to Pseudomonas syringae pv. actinidiae biovar 3. J Gen Plant Pathol, 2018, 84: 399-406

[27]

Kumar R, Bohra A, Pandey AK, Pandey MK, Kumar A. Metabolomics for plant improvement: status and prospects. Front Plant Sci, 2017, 8: 1302

[28]

Langfelder P, Horvath S. WGCNA: an R package for weighted correlation network analysis. BMC Bioinformatics, 2008, 9: 559

[29]

Li B, Li X, Wang Y, Liu X, Li K, Li R, et al.. KPGD: a kiwifruit pangenome database for comprehensive mining of genetic diversity in the genus Actinidia. Plant Commun, 2025, 6 Article ID: 101373

[30]

Li H, Durbin R. Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics, 2009, 25: 1754-1760

[31]

Liu Y, Li D, Zhang Q, Song C, Zhong C, Zhang X, et al.. Rapid radiations of both kiwifruit hybrid lineages and their parents shed light on a two-layer mode of species diversification. New Phytol, 2017, 215: 877-890

[32]

Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol, 2014, 15: 550

[33]

McKenna A, Hanna M, Banks E, Sivachenko A, Cibulskis K, Kernytsky A, et al.. The Genome Analysis Toolkit: a MapReduce framework for analyzing next-generation DNA sequencing data. Genome Res, 2010, 20: 1297-1303

[34]

Miller SA, Smith GS, Cookson JH. The effects of transient waterlogging on different species of Actinidia. Acta Hortic. 1997;444:323–8. https://doi.org/10.17660/ActaHortic.1997.444.50.

[35]

Nachtweide S, Stanke M. Multi-genome annotation with AUGUSTUS. Methods Mol Biol, 2019, 1962: 139-160

[36]

Nazir F, Fariduddin Q, Hussain A, Khan TA. Brassinosteroid and hydrogen peroxide improve photosynthetic machinery, stomatal movement, root morphology and cell viability and reduce Cu-triggered oxidative burst in tomato. Ecotoxicol Environ Saf, 2021, 207: 111081

[37]

Pan J, Sharif R, Xu X, Chen X. Mechanisms of waterlogging tolerance in plants: research progress and prospects. Front Plant Sci, 2021, 11: 627331

[38]

Pilkington SM, Crowhurst R, Hilario E, Nardozza S, Fraser L, Peng Y, et al.. A manually annotated Actinidia chinensis var. chinensis (kiwifruit) genome highlights the challenges associated with draft genomes and gene prediction in plants. BMC Genomics, 2018, 19: 257

[39]

Price MN, Dehal PS, Arkin AP. FastTree: computing large minimum evolution trees with profiles instead of a distance matrix. Mol Biol Evol, 2009, 26: 1641-1650

[40]

Sanz V, López-Hortas L, Torres MD, Domínguez H. Trends in kiwifruit and byproducts valorization. Trends Food Sci Technol, 2021, 107: 401-414

[41]

Simao FA, Waterhouse RM, Ioannidis P, Kriventseva EV, Zdobnov EM. BUSCO: assessing genome assembly and annotation completeness with single-copy orthologs. Bioinformatics, 2015, 31: 3210-3212

[42]

Slater GSC, Birney E. Automated generation of heuristics for biological sequence comparison. BMC Bioinformatics, 2005, 6 Article ID: 31

[43]

Suezawa K, Fukuda T, Mizutani R, Yamashita T, Otani M, Abe M, et al. Field performance of tetraploid Actinidia chinensis 'Sanuki Gold' on Actinidia rufa rootstocks. Acta Hortic. 2018;1218:413–8. https://doi.org/10.17660/ActaHortic.2018.1218.57.

[44]

Suleria HAR, Barrow CJ, Dunshea FR. Screening and characterization of phenolic compounds and their antioxidant capacity in different fruit peels. Foods, 2020, 9: 1206

[45]

Tang H, Krishnakumar V, Zeng X, Xu Z, Taranto A, Lomas JS, et al.. JCVI: a versatile toolkit for comparative genomics analysis. Imeta, 2024, 3: e211

[46]

Tang W, Sun X, Yue J, Tang X, Jiao C, Yang Y, et al.. Chromosome-scale genome assembly of kiwifruit Actinidia eriantha with single-molecule sequencing and chromatin interaction mapping. Gigascience, 2019, 8: giz027

[47]

Tarailo-Graovac M, Chen N. Using RepeatMasker to identify repetitive elements in genomic sequences. Curr Protoc Bioinformatics, 2009, 25: 4.10.1-4.10.14

[48]

Terhorst J, Kamm JA, Song YS. Robust and scalable inference of population history from hundreds of unphased whole genomes. Nat Genet, 2017, 49: 303-309

[49]

Vollering J, Halvorsen R, Mazzoni S. The MIAmaxent R package: variable transformation and model selection for species distribution models. Ecol Evol, 2019, 9: 12051-12068

[50]

Wang S, Qiu Y, Zhu F. Kiwifruit (Actinidia spp.): a review of chemical diversity and biological activities. Food Chem, 2021, 350: 128469

[51]

Wang X, Cao M, Li H, Liu Y, Fan S, Zhang N, et al.. Strategies and prospects for melatonin to alleviate abiotic stress in horticultural plants. Hortic Plant J, 2024, 10: 601-614

[52]

Wang Y, Dong J, Yuan X, Zheng H, Zhong C, Zhang Q. Cracking the kiwifruit code: how cultivar and origin co-create premium quality and consumer appeal. Future Foods, 2025, 11: 100651

[53]

Wang Y, Dong M, Wu Y, Zhang F, Ren W, Lin Y, et al.. Telomere-to-telomere and haplotype-resolved genome of the kiwifruit Actinidia eriantha. Mol Hortic, 2023, 3: 4

[54]

Wu H, Ma T, Kang M, Ai F, Zhang J, Dong G, et al.. A high-quality Actinidia chinensis (kiwifruit) genome. Hortic Res, 2019, 6: 117

[55]

Wu H, Yang W, Dong G, Hu Q, Li D, Liu J. Construction of the super pan-genome for the genus Actinidia reveals structural variations linked to phenotypic diversity. Hortic Res, 2025, 12: uhaf067

[56]

Wu L, Dong J, Zheng H, Wang Y, Han F, Yang S, et al.. Transcription factor AcNAC056 and AcMADS1 regulate gibberellin 2-oxidase gene AcGA2ox1 induced parthenocarpy in Actinidia eriantha. Int J Biol Macromol, 2025, 322: 146917

[57]

Wu L, Dong J, Zheng H, Wang Y, Qiu D, Atak A, et al.. Effects of plant growth regulator (2,4-D) treatments on fruit development and chemical content in the kiwifruit plant (Actinidia chinensis). BMC Plant Biol, 2025, 25: 1073

[58]

Xia H, Deng H, Li M, Xie Y, Lin L, Zhang H, et al.. Chromosome-scale genome assembly of a natural diploid kiwifruit (Actinidia chinensis var. deliciosa). Sci Data, 2023, 10: 92

[59]

Xu L, Miao T, Cui Z, Varkonyi-Gasic E, Zhang A, Wang L, et al.. Synteny-based comparative pan-genome reveals a male-specific FT gene underlying flowering time dimorphism in kiwifruit. Plant J, 2026, 125: e70664

[60]

Xu P, Fang S, Chen H, Cai W. The brassinosteroid-responsive xyloglucan endotransglucosylase/hydrolase 19 (XTH19) and XTH23 genes are involved in lateral root development under salt stress in Arabidopsis. Plant J, 2020, 104: 59-75

[61]

Xu W, Campbell P, Vargheese AK, Braam J. The Arabidopsis XET-related gene family: environmental and hormonal regulation of expression. Plant J, 1996, 9: 879-889

[62]

Xu Z, Wang H. LTR_FINDER: an efficient tool for the prediction of full-length LTR retrotransposons. Nucleic Acids Res, 2007, 35: W265-W268

[63]

Yang L, Fang S, Liu L, Zhao L, Chen W, Li X, et al.. WRKY transcription factors: hubs for regulating plant growth and stress responses. J Integr Plant Biol, 2025, 67: 488-509

[64]

Yang Z. PAML 4: phylogenetic analysis by maximum likelihood. PAML4: Phylogenetic Analysis by Maximum Likelihood Mol Biol Evol, 2007, 24: 1586-1591

[65]

Yao X, Wang S, Wang Z, Li D, Jiang Q, Zhang Q, et al.. The genome sequencing and comparative analysis of a wild kiwifruit Actinidia eriantha. Mol Hortic, 2022, 2: 13

[66]

Yu X, Qin M, Qu M, Jiang Q, Guo S, Chen Z, et al.. Genomic analyses reveal dead-end hybridization between two deeply divergent kiwifruit species rather than homoploid hybrid speciation. Plant J, 2023, 115: 1528-1543

[67]

Yuan X, Zheng H, Fan J, Liu F, Li J, Zhong C, et al.. Comparative study on physicochemical and nutritional qualities of kiwifruit varieties. Foods, 2022, 12: 108

[68]

Yue J, Chen Q, Wang Y, Zhang L, Ye C, Wang X, et al.. Telomere-to-telomere and gap-free reference genome assembly of the kiwifruit Actinidia chinensis. Hortic Res, 2023, 10: uhac264

[69]

Zhang C, He M, Jiang Z, Liu L, Pu J, Zhang W, et al.. The xyloglucan endotransglucosylase/hydrolase gene XTH22/TCH4 regulates plant growth by disrupting the cell wall homeostasis in Arabidopsis under boron deficiency. Int J Mol Sci, 2022, 23: 1250

[70]

Zhang H, Li G, Yan C, Zhang X, Cao N, Le M, et al.. Elucidating the molecular responses to waterlogging stress in Cucumis melo by comparative transcriptome profiling. Horticulturae, 2022, 8: 891

[71]

Zhang J, Huang S, Mo Z, Xuan J, Jia X, Wang G, et al.. De novo transcriptome sequencing and comparative analysis of differentially expressed genes in kiwifruit under waterlogging stress. Mol Breed, 2015, 35: 1-12

[72]

Zhang L, Tang Z, Zheng H, Zhong C, Zhang Q. Comprehensive analysis of metabolome and transcriptome in fruits and roots of kiwifruit. Int J Mol Sci, 2023, 24: 1299

[73]

Zhang Q, Liu C, Liu Y, VanBuren R, Yao X, Zhong C, et al.. High-density interspecific genetic maps of kiwifruit and the identification of sex-specific markers. DNA Res, 2015, 22: 367-375

[74]

Zhang Z, Xiao J, Wu J, Zhang H, Liu G, Wang X, Dai L. ParaAT: a parallel tool for constructing multiple protein-coding DNA alignments. Biochem Biophys Res Commun, 2012, 419: 779-781

[75]

Zheng L, Gao C, Zhao C, Zhang L, Han M, An N, et al.. Effects of brassinosteroid associated with auxin and gibberellin on apple tree growth and gene expression patterns. Hortic Plant J, 2019, 5: 93-108

[76]

Zheng X, Zhou J, Tan DX, Wang N, Wang L, Shan D, et al.. Melatonin improves waterlogging tolerance of Malus baccata (Linn.) Borkh. seedlings by maintaining aerobic respiration, photosynthesis and ROS migration. Front Plant Sci, 2017, 8: 483

[77]

Zhong C, Li D, Zhang Q, Li L, Huang W, Han, F. Identification, creation and utilization of the germplasm resources of kiwifruit. Acta Hortic. 2024;1401:17–24. https://doi.org/10.17660/ActaHortic.2024.1401.3.

[78]

Zhou Y, Zhang Z, Zhao X, Liu L, Tang Q, Fu J, et al.. Receptor-like cytoplasmic kinase STK confers salt tolerance in rice. Rice, 2023, 16: 21

[79]

Zimin AV, Salzberg SL. The genome polishing tool POLCA makes fast and accurate corrections in genome assemblies. PLoS Comput Biol, 2020, 16: e1007981

Funding

the earmarked fund for CARS(CARS26)

the National Key R&D Program of China(2022YFD1400200)

the Scientific and Technological Innovation Project of Hainan Provincial Investment and Holding Group Co., Ltd.(HKKJ2025JBGS09)

the Guangxi Science and Technology Major Project(Guike AA23023008)

Hubei Provincial Technological Innovation Program(2024BBB020)

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